The present invention relates to inhibitors of p38, a mammalian protein kinase involved cell proliferation, cell death and response to extracellular stimuli. The invention also relates to methods for producing these inhibitors. The invention also provides pharmaceutical compositions comprising the inhibitors of the invention and methods of utilizing those compositions in the treatment and prevention of various disorders.
Protein kinases are involved in various cellular responses to extracellular signals. Recently, a family of mitogen-activated protein kinases (MAPK) have been discovered. Members of this family are Ser/Thr kinases that activate their substrates by phosphorylation [B. Stein et al., Ann. Rep. Med. Chem., 31, pp. 289-98 (1996)]. MAPKs are themselves activated by a variety of signals including growth factors, cytokines, UV radiation, and stress-inducing agents.
One particularly interesting MAPK is p38. p38, also known as cytokine suppressive anti-inflammatory drug binding protein (CSBP) and RK, was isolated from murine pre-B cells that were transfected with the lipopolysaccharide (LPS) receptor CD14 and induced with LPS. p38 has since been isolated and sequenced, as has the cDNA encoding it in humans and mouse. Activation of p38 has been observed in cells stimulated by stresses, such as treatment of lipopolysaccharides (LPS), UV, anisomycin, or osmotic shock, and by cytokines, such as IL-1 and TNF.
Inhibition of p38 kinase leads to a blockade on the production of both IL-1 and TNF. IL-1 and TNF stimulate the production of other proinflammatory cytokines such as IL-6 and IL-8 and have been implicated in acute and chronic inflammatory diseases and in post-menopausal osteoporosis [R. B. Kimble et al., Endocrinol., 136, pp. 3054-61 (1995)].
Based upon this finding it is believed that p38, along with other MAPKs, have a role in mediating cellular response to inflammatory stimuli, such as leukocyte accumulation, macrophage/monocyte activation, tissue resorption, fever, acute phase responses and neutrophilia. In addition, MAPKs, such as p38, have been implicated in cancer, thrombin-induced platelet aggregation, immunodeficiency disorders, autoimmune diseases, cell death, allergies, osteoporosis and neurodegenerative disorders. Inhibitors of p38 have also been implicated in the area of pain management through inhibition of prostaglandin endoperoxide synthase-2 induction. Other diseases associated with Il-1, IL-6, IL-8 or TNF overproduction are set forth in WO 96/21654.
Others have already begun trying to develop drugs that specifically inhibit MAPKs. For example, PCT publication WO 95/31451 describes pyrazole compounds that inhibit MAPKs, and in particular p38. However, the efficacy of these inhibitors in vivo is still being investigated.
Accordingly, there is still a great need to develop other potent, p38-specific inhibitors that are useful in treating various conditions associated with p38 activation.
The present invention solves this problem by providing compounds which demonstrate strong and specific inhibition of p38.
These compounds have the general formula: 
wherein each of Q1 and Q2 are independently selected from 5-6 membered aromatic carbocyclic or heterocyclic ring systems, or 8-10 membered bicyclic ring systems comprising aromatic carbocyclic rings, aromatic heterocyclic rings or a combination of an aromatic carbocyclic ring and an aromatic heterocyclic ring.
The rings that make up Q1 are substituted with 1 to 4 substituents, each of which is independently selected from halo; C1-C3 alkyl optionally substituted with NRxe2x80x22, ORxe2x80x2, CO2Rxe2x80x2 or CONRxe2x80x22; Oxe2x80x94(C1-C3)-alkyl optionally substituted with NRxe2x80x22, ORxe2x80x2, CO2Rxe2x80x2 or CONRxe2x80x22; NRxe2x80x22; OCF3; CF3; NO2; CO2Rxe2x80x2; CONHRxe2x80x2; SRxe2x80x2; S(O2)N(Rxe2x80x2)2; SCF3; CN; N(Rxe2x80x2)C(O)R4; N(Rxe2x80x2)C(O)OR4; N(Rxe2x80x2)C(O)C(O)R4; N(Rxe2x80x2)S(O2)R4; N(Rxe2x80x2)R4; N(R4)2; OR4; OC(O)R4; OP(O)3H2; or Nxe2x95x90CHxe2x80x94N(Rxe2x80x2)2.
The rings that make up Q2 are optionally substituted with up to 4 substituents, each of which is independently selected from halo; C1-C3 straight or branched alkyl optionally substituted with NRxe2x80x22, ORxe2x80x2, CO2Rxe2x80x2, S(O2)N(Rxe2x80x2)2, Nxe2x95x90CHxe2x80x94N(Rxe2x80x2)2, R3, or CONRxe2x80x22; Oxe2x80x94(C1-C3)-alkyl; Oxe2x80x94(C1-C3)-alkyl optionally substituted with NRxe2x80x22, ORxe2x80x2, CO2Rxe2x80x2, S(O2)N(Rxe2x80x2)2, Nxe2x95x90CHxe2x80x94N(Rxe2x80x2)2, R3, or CONRxe2x80x22; NRxe2x80x22; OCF3; CF3; NO2; CO2Rxe2x80x2; CONHRxe2x80x2; R3; OR3; NHR3; SR3; C(O)R3; C(O)N(Rxe2x80x2)R3; C(O)OR3; SRxe2x80x2; S(O2) N(Rxe2x80x2)2; SCF3; Nxe2x95x90CHxe2x80x94N(Rxe2x80x2)2, or CN.
Rxe2x80x2 is selected from hydrogen, (C1-C3)-alkyl; (C2-C3)-alkenyl or alkynyl; phenyl or phenyl substituted with 1 to 3 substituents independently selected from halo, methoxy, cyano, nitro, amino, hydroxy, methyl or ethyl.
R3 is selected from 5-6 membered aromatic carbocyclic or heterocyclic ring systems.
R4 is (C1-C4)-alkyl optionally substituted with N(Rxe2x80x2)2, ORxe2x80x2, CO2Rxe2x80x2, CON(Rxe2x80x2)2, or SO2N(R2)2; or a 5-6 membered carbocyclic or heterocyclic ring system optionally substituted with N(Rxe2x80x2)2, ORxe2x80x2, CO2Rxe2x80x2, CON(Rxe2x80x2)2, or SO2N(R2)2.
X is selected from xe2x80x94Sxe2x80x94, xe2x80x94Oxe2x80x94, xe2x80x94S(O2)xe2x80x94, xe2x80x94S(O)xe2x80x94, xe2x80x94S(O2)xe2x80x94N(R2)xe2x80x94, xe2x80x94N(R2)xe2x80x94S(O2)xe2x80x94, xe2x80x94N(R)xe2x80x94C(O)Oxe2x80x94, xe2x80x94Oxe2x80x94C(O)xe2x80x94N(R2), xe2x80x94C(O)xe2x80x94, xe2x80x94C(O)Oxe2x80x94, xe2x80x94Oxe2x80x94C(O)xe2x80x94, xe2x80x94C(O)xe2x80x94N(R2)xe2x80x94, xe2x80x94N(R2)xe2x80x94C(O)xe2x80x94, xe2x80x94N(R2)xe2x80x94, xe2x80x94C(R2)2xe2x80x94, or xe2x80x94C(OR2)2.
Each R is independently selected from hydrogen, xe2x80x94R2, xe2x80x94N(R2)2, xe2x80x94OR2, SR2, xe2x80x94C(O)xe2x80x94N(R2)2, xe2x80x94S(O2)xe2x80x94N(R2)2, or xe2x80x94C(O)xe2x80x94OR2, wherein two adjacent R are optionally bound to one another and, together with each Y to which they are respectively bound, form a 4-8 membered carbocyclic or heterocyclic ring;
R2 is selected from hydrogen, (C1-C3)-alkyl, or (C1-C3)-alkenyl; each optionally substituted with xe2x80x94N(Rxe2x80x2)2, xe2x80x94ORxe2x80x2, SRxe2x80x2, xe2x80x94C(O)xe2x80x94N(Rxe2x80x2)2, xe2x80x94S(O2)xe2x80x94N(Rxe2x80x2)2, xe2x80x94C(O)xe2x80x94ORxe2x80x2, or R3.
Y is N or C;
A, if present, is N or CRxe2x80x2;
n is 0 or 1;
R1 is selected from hydrogen, (C1-C3)-alkyl, OH, or Oxe2x80x94(C1-C3)-alkyl.
In another embodiment, the invention provides pharmaceutical compositions comprising the p38 inhibitors of this invention. These compositions may be utilized in methods for treating or preventing a variety of disorders, such as cancer, inflammatory diseases, autoimmune diseases, destructive bone disorders, proliferative disorders, infectious diseases, viral diseases and neurodegenerative diseases. These compositions are also useful in methods for preventing cell death and hyperplasia and therefore may be used to treat or prevent reperfusion/ischemia in stroke, heart attacks, organ hypoxia. The compositions are also useful in methods for preventing thrombin-induced platelet aggregation. Each of these above-described methods is also part of the present invention.
The present invention provides inhibitors of p38 having the general formula: 
wherein each of Q1 and Q2 are independently selected from 5-6 membered aromatic carbocyclic or heterocyclic ring systems, or 8-10 membered bicyclic ring systems comprising aromatic carbocyclic rings, aromatic heterocyclic rings or a combination of an aromatic carbocyclic ring and an aromatic heterocyclic ring.
The rings that make up Q1 are substituted with 1 to 4 substituents, each of which is independently selected from halo; C1-C3 alkyl optionally substituted with NRxe2x80x22, ORxe2x80x2, CO2Rxe2x80x2 or CONRxe2x80x22; Oxe2x80x94(C1-C3)-alkyl optionally substituted with NRxe2x80x22, ORxe2x80x2, CO2Rxe2x80x2 or CONRxe2x80x22; NRxe2x80x22; OCF3; CF3; NO2; CO2Rxe2x80x2; CONHRxe2x80x2; SRxe2x80x2; S(O2)N(Rxe2x80x2)2; SCF3; CN; N(Rxe2x80x2)C(O)R4; N(Rxe2x80x2)C(O)OR4; N(Rxe2x80x2)C(O)C(O)R4; N(Rxe2x80x2)S(O2)R4; N(Rxe2x80x2)R4; N(R4)2; OR4; OC(O)R4; OP(O)3H2; or Nxe2x95x90CHxe2x80x94N(Rxe2x80x2)2.
The rings that make up Q2 are optionally substituted with up to 4 substituents, each of which is independently selected from halo; C1-C3 straight or branched alkyl optionally substituted with NRxe2x80x22, ORxe2x80x2, CO2Rxe2x80x2, S(O2)N(Rxe2x80x2)2, Nxe2x95x90CHxe2x80x94N(Rxe2x80x2)2, R3, or CONRxe2x80x22; Oxe2x80x94(C1-C3)-alkyl; Oxe2x80x94(C1-C3)-alkyl optionally substituted with NRxe2x80x22, ORxe2x80x2, CO2Rxe2x80x2, S(O2)N(Rxe2x80x2)2, Nxe2x95x90CHxe2x80x94N(Rxe2x80x2)2, R3, or CONRxe2x80x22; NRxe2x80x22; OCF3; CF3; NO2; CO2Rxe2x80x2; CONHRxe2x80x2; R3; OR3; NHR3; SR3; C(O)R3; C(O)N(Rxe2x80x2)R3; C(O)OR3; SRxe2x80x2; S(O2)N(Rxe2x80x2)2; SCF3; Nxe2x95x90CHxe2x80x94N(Rxe2x80x2)2, or CN.
Rxe2x80x2 is selected from hydrogen, (C1-C3)-alkyl; (C2-C3)-alkenyl or alkynyl; phenyl or phenyl substituted with 1 to 3 substituents independently selected from halo, methoxy, cyano, nitro, amino, hydroxy, methyl or ethyl.
R3 is selected from 5-6 membered aromatic carbocyclic or heterocyclic ring systems.
R4 is (C1-C4)-alkyl optionally substituted with N(Rxe2x80x2)2, ORxe2x80x2, CO2Rxe2x80x2, CON(Rxe2x80x2)2, or SO2N(R2)2; or a 5-6 membered carbocyclic or heterocyclic ring system optionally substituted with N(Rxe2x80x2)2, ORxe2x80x2, CO2Rxe2x80x2, CON(Rxe2x80x2)2, or SO2N(R2)2.
X is selected from xe2x80x94Sxe2x80x94, xe2x80x94Oxe2x80x94, xe2x80x94S(O2)xe2x80x94, xe2x80x94S(O)xe2x80x94, xe2x80x94S(O2)xe2x80x94N(R2)xe2x80x94, xe2x80x94N(R2)xe2x80x94S(O2)xe2x80x94, xe2x80x94N(R2)xe2x80x94C(O)Oxe2x80x94, xe2x80x94Oxe2x80x94C(O)xe2x80x94N(R2), xe2x80x94C(O)xe2x80x94, xe2x80x94C(O)Oxe2x80x94, xe2x80x94Oxe2x80x94C(O)xe2x80x94, xe2x80x94C(O)xe2x80x94N(R2)xe2x80x94, xe2x80x94N(R2)xe2x80x94C(O)xe2x80x94, xe2x80x94N(R2)xe2x80x94, xe2x80x94C(R2)2xe2x80x94, or xe2x80x94C(OR2)2xe2x80x94.
Each R is independently selected from hydrogen, xe2x80x94R2, xe2x80x94N(R2)2, xe2x80x94OR2, SR2, xe2x80x94C(O)xe2x80x94N(R2)2, xe2x80x94S(O2)xe2x80x94N(R2)2, or xe2x80x94C(O)xe2x80x94OR2, wherein two adjacent R are optionally bound to one another and, together with each Y to which they are respectively bound, form a 4-8 membered carbocyclic or heterocyclic ring;
When the two R components form a ring together with the Y components to which they are respectively bound, it will obvious to those skilled in the art that a terminal hydrogen from each unfused R component will be lost. For example, if a ring structure is formed by binding those two R components together, one being xe2x80x94NHxe2x80x94CH3 and the other being xe2x80x94CH2xe2x80x94CH3, one terminal hydrogen on each R component (indicated in bold) will be lost. Therefore, the resulting portion of the ring structure will have the formula xe2x80x94NHxe2x80x94CH2xe2x80x94CH2xe2x80x94CH2xe2x80x94.
R2 is selected from hydrogen, (C1-C3)-alkyl, or (C1-C3)-alkenyl; each optionally substituted with xe2x80x94N(Rxe2x80x2)2, xe2x80x94ORxe2x80x2, SRxe2x80x2, xe2x80x94C(O)xe2x80x94N(Rxe2x80x2)2, xe2x80x94S(O2)xe2x80x94N(Rxe2x80x2)2, xe2x80x94C(O)xe2x80x94ORxe2x80x2, or R3.
Y is N or C;
A, if present, is N or CRxe2x80x2;
n is 0 or 1;
R1 is selected from hydrogen, (C1-C3)-alkyl, OH, or Oxe2x80x94(C1-C3)-alkyl. It will be apparent to those of skill in the art that if R1 is OH, the resulting inhibitor may tautomerize resulting in a compound of the formula: 
which are also p38 inhibitors of this invention.
According to another preferred embodiment, Q1 is selected from phenyl or pyridyl containing 1 to 3 substituents, wherein at least one of said substituents is in the ortho position and said substituents are independently selected from chloro, fluoro, bromo, xe2x80x94CH3, xe2x80x94OCH3, xe2x80x94OH, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94O(CH2)2CH3, NH2, 3,4-methylenedioxy, xe2x80x94N(CH3)2, xe2x80x94NHxe2x80x94S(O)2-phenyl, xe2x80x94NHxe2x80x94C(O)Oxe2x80x94CH2-4-pyridine, xe2x80x94NHxe2x80x94C(O)CH2-morpholine, xe2x80x94NHxe2x80x94C(O)CH2xe2x80x94N(CH3)2, xe2x80x94NHxe2x80x94C(O)CH2-piperazine, xe2x80x94NHxe2x80x94C(O)CH2-pyrrolidine, xe2x80x94NHxe2x80x94C(O)C(O)-morpholine, xe2x80x94NHxe2x80x94C(O)C(O)-piperazine, xe2x80x94NHxe2x80x94C(O)C(O)-pyrrolidine, xe2x80x94Oxe2x80x94C(O)CH2xe2x80x94N(CH3)2, or xe2x80x94Oxe2x80x94(CH2)2xe2x80x94N(CH3)2.
Even more preferred are phenyl or pyridyl containing at least 2 of the above-indicated substituents both being in the ortho position.
Some specific examples of preferred Q1 are: 
Most preferably, Q1 is selected from 2-fluoro-6-trifluoromethylphenyl, 2,6-difluorophenyl, 2,6-dichlorophenyl, 2-chloro-4-hydroxyphenyl, 2-chloro-4-aminophenyl, 2,6-dichloro-4-aminophenyl, 2,6-dichloro-3-aminophenyl, 2,6-dimethyl-4-hydroxyphenyl, 2-methoxy-3,5-dichloro-4-pyridyl, 2-chloro-4,5 methylenedioxy phenyl, or 2-chloro-4-(N-2-morpholino-acetamido)phenyl.
According to a preferred embodiment, Q2 is phenyl or pyridyl containing 0 to 3 substituents, wherein each substituent is independently selected from chloro, fluoro, bromo, methyl, ethyl, isopropyl, xe2x80x94OCH3, xe2x80x94OH, xe2x80x94NH2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94SCH3, xe2x80x94OCH3, xe2x80x94C(O)OH, xe2x80x94C(O)OCH3, xe2x80x94CH2NH2, xe2x80x94N(CH3)2, xe2x80x94CH2-pyrrolidine and xe2x80x94CH2OH.
Some specific examples of preferred Q2 are: 
unsubstituted 2-pyridyl or unsubstituted phenyl.
Most preferred are compounds wherein Q2 is selected from phenyl, 2-isopropylphenyl, 3,4-dimethylphenyl, 2-ethylphenyl, 3-fluorophenyl, 2-methylphenyl, 3-chloro-4-fluorophenyl, 3-chlorophenyl, 2-carbomethoxylphenyl, 2-carboxyphenyl, 2-methyl-4-chlorophenyl, 2-bromophenyl, 2-pyridyl, 2-methylenehydroxyphenyl, 4-fluorophenyl, 2-methyl-4-fluorophenyl, 2-chloro-4-fluorphenyl, 2,4-difluorophenyl, 2-hydroxy-4-fluorphenyl or 2-methylenehydroxy-4-fluorophenyl.
According to yet another preferred embodiment, X is xe2x80x94Sxe2x80x94, xe2x80x94Oxe2x80x94, xe2x80x94S(O2)xe2x80x94, xe2x80x94S(O)xe2x80x94, xe2x80x94NRxe2x80x94, xe2x80x94C(R2)xe2x80x94 or xe2x80x94C(O)xe2x80x94. Most preferably, X is S.
According to another preferred embodiment, n is 1 and A is N.
According to another preferred embodiment, each Y is C.
According an even more preferred embodiment, each Y is C and the R attached to those Y components is selected from hydrogen or methyl.
Some specific inhibitors of this invention are set forth in the table below.
According to another embodiment, the present invention provides methods of producing inhibitors of p38 of the formula (Ia) depicted above. These methods involve reacting a compound of formula II: 
wherein each of the variables in the above formula are the same as defined above for the inhibitors of this invention, with a leaving group reagent of formula IIa: 
wherein Rxe2x80x2 is as defined above, or a leaving group reagent of formula IIb: 
wherein each of L1, L2, and L3 independently represents a leaving group.
The leaving group reagent used in this reaction is added in excess, either neat or with a co-solvent, such as toluene. The reaction is carried out at a temperature of between 25xc2x0 C. and 150xc2x0 C.
Leaving group reagents of formula IIa that are useful in producing the p38 inhibitors of this invention include dimethylformamide dimethylacetal, dimethylacetamide dimethylacetal, trimethyl orthoformate, dimethylformamide diethylacetal and other related reagents. Preferably the leaving group reagents of formula IIa used to produce the inhibitors of this invention is dimethylformamide dimethylacetal.
Leaving group reagents of formula IIb that are useful in producing the p38 inhibitors of this invention include phosgene, carbonyldiimidazole, diethyl carbonate and triphosgene.
More preferred methods of producing the compounds of this invention utilize compounds of formula II wherein each of the variables are defined in terms of the more preferred and most preferred choices as set forth above for the compounds of this invention.
Because the source of R1 is the leaving group reagent (Cxe2x80x94Rxe2x80x2 or Cxe2x95x90O), its identity is, of course, dependent on the structure of that reagent. Therefore, in compounds where R1 is OH, the reagent used must be IIb. Similarly, when R1 is H or (C1-C3)-alkyl, the reagent used must be IIa. In order to generate inhibitors wherein R1 is Oxe2x80x94(C1-C3)-alkyl, a compound wherein R1 is OH is first generated, followed by alkylation of that hydroxy by standard techniques, such as treatment with Na hydride in DMF, methyl iodide and ethyl iodide.
The immediate precursors to the inhibitors of this invention of formula Ia (i.e., compounds of Formula II) may themselves be synthesized by either of the synthesis schemes depicted below: 
In Scheme 1, the order of steps 1) and 2) can be reversed. Also, the starting nitrile may be replaced by a corresponding acid or by an ester. Alternatively, other well-known latent carboxyl or carboxamide moieties may be used in place of the nitrile (see scheme 2). Variations such as carboxylic acids, carboxylic esters, oxazolines or oxizolidinones may be incorporated into this scheme by utilizing subsequent deprotection and functionalization methods which are well known in the art
The base used in the first step of Scheme 1 (and in Scheme 2, below) is selected from sodium hydride, sodium amide, LDA, lithium hexamethyldisilazide, sodium hexamethyldisilazide or any number of other non-nucleophilic bases that will deprotonate the position alpha to the nitrile.
Also, the addition of HX-Q2 in the single step depicted above may be substituted by two stepsxe2x80x94the addition of a protected or unprotected X derivative followed by the addition of a Q2 derivative in a subsequent step. 
In Scheme 2, Z is selected from COOH, COORxe2x80x2, CON(Rxe2x80x2)2, oxazoline, oxazolidinone or CN. Rxe2x80x2 is as defined above.
According to another embodiment, the present invention provides methods of producing inhibitors of p38 of the formula (Ib) depicted above. These methods involve reacting a compound of formula III: 
wherein each of the variables in the above formula are the same as defined above for the inhibitors of this invention, with a leaving group reagent of formula: 
as described above.
Two full synthesis schemes for the p38 inhibitors of formula (Ib) of this invention are depicted below. 
In scheme 3, a Q1 substituted derivative may be treated with a base such as sodium hydride, sodium amide, LDA, lithium hexamethyldisilazide, sodium hexamethyldisilazide or any number of other non-nucleophilic bases to deprotonate the position alpha to the Z group, which represents a masked amide moiety. Alternatively, Z is a carboxylic acid, carboxylic ester, oxazoline or oxazolidinone. The anion resulting from deprotonation is then contacted with a nitrogen bearing heterocyclic compound which contains two leaving groups, or latent leaving groups, in the presence of a Palladium catalyst. One example of such compound may be 2,6-dichloropyridine.
In step two, the Q2 ring moiety is introduced. This may be performed utilizing many reactions well known in the art which result in the production of biaryl compounds. One example may be the reaction of an aryl lithium compound with the pyridine intermediate produced in step 1. Alternatively, an arylmetallic compound such as an aryl stannane or an aryl boronic acid may be reacted with the aryl halide portion of the pyridine intermediate in the presence of a Pdo catalyst.
In step 3 the Z group is deprotected and/or functionalized to form the amide compound. When Z is a carboxylic acid, carboxylic ester, oxazoline or oxazolidinone, variations in deprotection and functionalization methods which are well known in the art are employed to produce the amide. Finally in step 4, the amide compound is cyclized to the final product utilizing reagents such as DMF acetal or similar reagents either neat or in an organic solvent. 
Scheme 4 is similar except that the a biaryl intermediate is first generated prior to reaction with the Q1 starting material.
According to another embodiment, the invention provides inhibitors of p38 similar to those of formulae Ia and Ib above, but wherein the Cxe2x95x90N in the ring bearing the Q1 substituent is reduced. These inhibitors have the formula: 
wherein A, Q1, Q2, R, Rxe2x80x2, X, Y and n are defined in the same manner as set forth for compounds of formulae Ia and Ib. These definitions hold for all embodiments of each of these variables (i.e., basic, preferred, more preferred and most preferred). R5 is selected from hydrogen, xe2x80x94CRxe2x80x22OH, xe2x80x94C(O)R4, xe2x80x94C(O)OR4, xe2x80x94CRxe2x80x22OPO3H2, xe2x80x94PO3H2, and salts of xe2x80x94PO3H2.
When R5 is not hydrogen, the resulting compounds are expected to be prodrug forms which should be cleaved in vivo to produce a compound wherein R5 is hydrogen.
According to other preferred embodiments, in compounds of formula Ic, A is preferably nitrogen, n is preferably 1, and X is preferably sulfur. In compounds of formula Ic or Id, Q1 and Q2 are preferably the same moieties indicated above for those variables in compounds of formulae Ia and Ib.
Compounds of formulae Ic and Id may be prepared directly from compounds of formulae Ia or Ib which contain a hydrogen, C1-C3 alkyl or C2-C3 alkenyl or alkynyl at the R1 position (e.g., where R1=Rxe2x80x2). The synthesis schemes for these compounds is depicted in Schemes 5 and 6, below. 
In these schemes, compounds of formula Ia or Ib are reduced by reaction with an excess of diisobutylaluminum hydride, or equivalent reagent to yield the ring reduced compounds of formula Ic or Id, respectively.
The addition of an R5 component other than hydrogen onto the ring nitrogen is achieved by reacting the formula Ic or Id compounds indicated above with the appropriate reagent(s). Examples of such modifications are provided in the Example section below.
Some specific inhibitors of this invention of formula Ic are set forth in the table below.
According to yet another embodiment, the invention provides p38 inhibitors of the formulae: 
wherein A, Q1, Q2, R, X, Y and n are defined in the same manner as set forth for compounds of formulae Ia and Ib. These definitions hold for all embodiments of each of these variables (i.e., basic, preferred, more preferred and most preferred). More preferably, in compounds of formula Ie, Q2 is unsubstituted phenyl.
Q3 is a 5-6 membered aromatic carbocyclic or heterocyclic ring system, or an 8-10 membered bicyclic ring system comprising aromatic carbocyclic rings, aromatic heterocyclic rings or a combination of an aromatic carbocyclic ring and an aromatic heterocyclic ring. The rings of Q3 are substituted with 1 to 4 substituents, each of which is independently selected from halo; C1-C3 alkyl optionally substituted with NRxe2x80x22, ORxe2x80x2, CO2Rxe2x80x2 or CONRxe2x80x22; Oxe2x80x94(C1-C3)-alkyl optionally substituted with NRxe2x80x22, ORxe2x80x2, CO2Rxe2x80x2 or CONRxe2x80x22; NRxe2x80x22; OCF3; CF3; NO2; CO2Rxe2x80x2; CONRxe2x80x2; SRxe2x80x2; S(O2)N(Rxe2x80x2)2; SCF3; CN; N(Rxe2x80x2)C(O)R4; N(Rxe2x80x2)C(O)OR4; N(Rxe2x80x2)C(O)C(O)R4; N(Rxe2x80x2)S(O2)R4; N(Rxe2x80x2)R4; N(R4)2; OR4; OC(O)R4; OP(O)3H2; or Nxe2x95x90Cxe2x80x94N(Rxe2x80x2)2.
According to one preferred embodiment, Q3 is substituted with 2 to 4 substituents, wherein at least one of said substituents is present in the ortho position relative to the point of attachment of Q3 to the rest of the inhibitor. When Q3 is a bicyclic ring, the 2 substituents in the ortho position are present on the ring that is closest (i.e., directly attached) to the rest of the inhibitor molecule. The other two optional substituents may be present on either ring. More preferably, both such ortho positions are occupied by one of said substituents.
According to another preferred embodiment, Q3 is a monocyclic carbocyclic ring, wherein each ortho substituent is independently selected from halo or methyl. According to another preferred embodiment, Q3 contains 1 or 2 additional substituents independently selected from NRxe2x80x22, ORxe2x80x2, CO2Rxe2x80x2 CN, N(Rxe2x80x2)C(O)R4; N(Rxe2x80x2)C(O)OR4; N(Rxe2x80x2)C(O)C(O)R4; N(Rxe2x80x2)S(O2)R4; N(Rxe2x80x2)R4; N(R4)2; OR4; OC(O)R4; OP(O)3H2; or Nxe2x95x90Cxe2x80x94N(Rxe2x80x2)2.
Preferably, Q3 is selected from any of the Q3 moieties present in the Ie compounds set forth in Table 3, below, or from any of the Q3 moieties present in the Ig compounds set forth in Table 4, below.
Those of skill will recognize compounds of formula Ie as being the direct precursors to certain of the formula Ia and formula Ic p38 inhibitors of this invention (i.e., those wherein Q1=Q3). Those of skill will also recognize that compounds of formula Ig are precursors to certain of the formula Ib and Id p38 inhibitors of this invention (i.e., those wherein Q1=Q3). Accordingly, the synthesis of formula Ie inhibitors is depicted above in Schemes 1 and 2, wherein Q1 is replaced by Q3. Similarly, the synthesis of formula Ig inhibitors is depicted above in Schemes 3 and 4, wherein Q1 is replaced by Q3.
The synthesis of formula If and formula Ih inhibitors is depicted below in Schemes 7 and 8. 
Scheme 8 depicts the synthesis of compounds of type Ih. For example, treating an initial dibromo derivative, such as 2,6 dibromopyridine, with an amine in the presence of a base such as sodium hydride yields the 2-amino-6-bromo derivative. Treatment of this intermediate with a phenylboronic acid analog (a Q2-boronic acid) such as phenyl boronic acid in the presence of a palladium catalyst gives the disubstituted derivative which can then be acylated to the final product. The order of the first two steps of this synthesis may be reversed.
Without being bound by theory, applicants believe that the diortho substitution in the Q3 ring of formula Ie and Ig inhibitors and the presence of a nitrogen directly attached to the Q1 ring in formula If and Ih inhibitors causes a xe2x80x9cflatteningxe2x80x9d of the compound that allows it to effectively inhibit p38.
A preferred formula Ie inhibitor of this invention is one wherein A is carbon, n is 1, X is sulfur, each Y is carbon, each R is hydrogen, Q3 is 2,6-dichlorophenyl and Q2 is phenyl, said compound being referred to as compound 201. A preferred formula Ig inhibitor of this invention is one wherein Q3 is 2,6-dichlorophenyl, Q2 is phenyl, each Y is carbon and each R is hydrogen. This compound is referred to herein as compound 202. Other preferred formula Ig compounds of this invention are those listed in Table 4, below.
Preferred Ih compounds of this invention are those depicted in Table 5, below. Other preferred Ih compounds are those wherein Q1 is phenyl independently substituted at the 2 and 6 positions by chloro or fluoro; each Y is carbon; each R is hydrogen; and Q2 is 2-methylphenyl, 4-fluorophenyl, 2,4-difluorophenyl, 2-methylenehydroxy-4-fluorophenyl, or 2-methyl-4-fluorophenyl.
Some specific inhibitors of formulae Ie, Ig and Ih are depicted in the tables below.
The activity of the p38 inhibitors of this invention may be assayed by in vitro, in vivo or in a cell line. In vitro assays include assays that determine inhibition of either the kinase activity or ATPase activity of activated p38. Alternate in vitro assays quantitate the ability of the inhibitor to bind to p38 and may be measured either by radiolabelling the inhibitor prior to binding, isolating the inhibitor/p38 complex and determining the amount of radiolabel bound, or by running a competition experiment where new inhibitors are incubated with p38 bound to known radioligands.
Cell culture assays of the inhibitory effect of the compounds of this invention may determine the amounts of TNF, IL-1, IL-6 or IL-8 produced in whole blood or cell fractions thereof in cells treated with inhibitor as compared to cells treated with negative controls. Level of these cytokines may be determined through the use of commercially available ELISAs.
An in vivo assay useful for determining the inhibitory activity of the p38 inhibitors of this invention are the suppression of hind paw edema in rats with Mycobacterium butyricum-induced adjuvant arthritis. This is described in J. C. Boehm et al., J. Med. Chem., 39, pp. 3929-37 (1996), the disclosure of which is herein incorporated by reference. The p38 inhibitors of this invention may also be assayed in animal models of arthritis, bone resorption, endotoxin shock and immune function, as described in A. M. Badger et al., J. Pharmacol. Experimental Therapeutics, 279, pp. 1453-61 (1996), the disclosure of which is herein incorporated by reference.
The p38 inhibitors or pharmaceutical salts thereof may be formulated into pharmaceutical compositions for administration to animals or humans. These pharmaceutical compositions, which comprise and amount of p38 inhibitor effective to treat or prevent a p38-mediated condition and a pharmaceutically acceptable carrier, are another embodiment of the present invention.
The term xe2x80x9cp38-mediated conditionxe2x80x9d, as used herein means any disease or other deleterious condition in which p38 is known to play a role. This includes, conditions which are known to be caused by IL-1, TNF, IL-6 or IL-8 overproduction. Such conditions include, without limitation, inflammatory diseases, autoimmune diseases, destructive bone disorders, proliferative disorders, infectious diseases, neurodegenerative diseases, allergies, reperfusion/ischemia in stroke, heart attacks, angiogenic disorders, organ hypoxia, vascular hyperplasia, cardiac hypertrophy, thrombin-induced platelet aggregation, and conditions associated with prostaglandin endoperoxidase synthase-2.
Inflammatory diseases which may be treated or prevented include, but are not limited to acute pancreatitis, chronic pancreatitis, asthma, allergies, and adult respiratory distress syndrome. Autoimmune diseases which may be treated or prevented include, but are not limited to, glomerulonephritis, rheumatoid arthritis, systemic lupus erythematosus, scleroderma, chronic thyroiditis, Graves"" disease, autoimmune gastritis, diabetes, autoimmune hemolytic anemia, autoimmune neutropenia, thrombocytopenia, atopic dermatitis, chronic active hepatitis, myasthenia gravis, multiple sclerosis, inflammatory bowel disease, ulcerative colitis, Crohn""s disease, psoriasis, or graft vs. host disease.
Destructive bone disorders which may be treated or prevented include, but are not limited to, osteoporosis, osteoarthritis and multiple myeloma-related bone disorder.
Proliferative diseases which may be treated or prevented include, but are not limited to, acute myelogenous leukemia, chronic myelogenous leukemia, metastatic melanoma, Kaposi""s sarcoma, and multiple myeloma.
Angiogenic disorders which may be treated or prevented include solid tumors, ocular neovasculization, infantile haemangiomas.
Infectious diseases which may be treated or prevented include, but are not limited to, sepsis, septic shock, and Shigellosis.
Viral diseases which may be treated or prevented include, but are not limited to, acute hepatitis infection (including hepatitis A, hepatitis B and hepatitis C), HIV infection and CMV retinitis.
Neurodegenerative diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, Alzheimer""s disease, Parkinson""s disease, cerebral ischemias or neurodegenerative disease caused by traumatic injury.
xe2x80x9cp38-mediated conditionsxe2x80x9d also include ischemia/reperfusion in stroke, heart attacks, myocardial ischemia, organ hypoxia, vascular hyperplasia, cardiac hypertrophy, and thrombin-induced platelet aggregation.
In addition, p38 inhibitors in this invention are also capable of inhibiting the expression of inducible pro-inflammatory proteins such as prostaglandin endoperoxide synthase-2 (PGHS-2), also referred to as cyclooxygenase-2 (COX-2). Therefore, other xe2x80x9cp38-mediated conditionsxe2x80x9d are edema, analgesia, fever and pain, such as neuromuscular pain, headache, cancer pain, dental pain and arthritis pain.
The diseases that may be treated or prevented by the p38 inhibitors of this invention may also be conveniently grouped by the cytokine (IL-1, TNF, IL-6, IL-8) that is believed to be responsible for the disease.
Thus, an IL-1-mediated disease or condition includes rheumatoid arthritis, osteoarthritis, stroke, endotoxemia and/or toxic shock syndrome, inflammatory reaction induced by endotoxin, inflammatory bowel disease, tuberculosis, atherosclerosis, muscle degeneration, cachexia, psoriatic arthritis, Reiter""s syndrome, gout, traumatic arthritis, rubella arthritis, acute synovitis, diabetes, pancreatic xcex2-cell disease and Alzheimer""s disease.
TNF-mediated disease or condition includes, rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthritis and other arthritic conditions, sepsis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, adult respiratory distress syndrome, cerebral malaria, chronic pulmonary inflammatory disease, silicosis, pulmonary sarcoisosis, bone resorption diseases, reperfusion injury, graft vs. host reaction, allograft rejections, fever and myalgias due to infection, cachexia secondary to infection, AIDS, ARC or malignancy, keloid formation, scar tissue formation, Crohn""s disease, ulcerative colitis or pyresis. TNF-mediated diseases also include viral infections, such as HIV, CMV, influenza and herpes; and veterinary viral infections, such as lentivirus infections, including, but not limited to equine infectious anemia virus, caprine arthritis virus, visna virus or maedi virus; or retrovirus infections, including feline immunodeficiency virus, bovine immunodeficiency virus, or canine immunodeficiency virus.
IL-8 mediated disease or condition includes diseases characterized by massive neutrophil infiltration, such as psoriasis, inflammatory bowel disease, asthma, cardiac and renal reperfusion injury, adult respiratory distress syndrome, thrombosis and glomerulonephritis.
In addition, the compounds of this invention may be used topically to treat or prevent conditions caused or exacerbated by IL-1 or TNF. Such conditions include inflamed joints, eczema, psoriasis, inflammatory skin conditions such as sunburn, inflammatory eye conditions such as conjunctivitis, pyresis, pain and other conditions associated with inflammation.
In addition to the compounds of this invention, pharmaceutically acceptable salts of the compounds of this invention may also be employed in compositions to treat or prevent the above-identified disorders.
Pharmaceutically acceptable salts of the compounds of this invention include those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acid salts include acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptanoate, glycerophosphate, glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oxalate, palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, salicylate, succinate, sulfate, tartrate, thiocyanate, tosylate and undecanoate. Other acids, such as oxalic, while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid addition salts. Salts derived from appropriate bases include alkali metal (e.g., sodium and potassium), alkaline earth metal (e.g., magnesium), ammonium and Nxe2x80x94(C1-4alkyl)4+ salts. This invention also envisions the quaternization of any basic nitrogen-containing groups of the compounds disclosed herein. Water or oil-soluble or dispersible products may be obtained by such quaternization.
Pharmaceutically acceptable carriers that may be used in these pharmaceutical compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.
The compositions of the present invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. The term xe2x80x9cparenteralxe2x80x9d as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally or intravenously.
Sterile injectable forms of the compositions of this invention may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer""s solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or di-glycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents which are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions. Other commonly used surfactants, such as Tweens, Spans and other emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.
The pharmaceutical compositions of this invention may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers which are commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.
Alternatively, the pharmaceutical compositions of this invention may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient which is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.
The pharmaceutical compositions of this invention may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.
Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches may also be used.
For topical applications, the pharmaceutical compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of this invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutical compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
For ophthalmic use, the pharmaceutical compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, preferably, as solutions in isotonic, pH adjusted sterile saline, either with or without a preservative such as benzylalkonium chloride. Alternatively, for ophthalmic uses, the pharmaceutical compositions may be formulated in an ointment such as petrolatum.
The pharmaceutical compositions of this invention may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.
The amount of p38 inhibitor that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated, the particular mode of administration. Preferably, the compositions should be formulated so that a dosage of between 0.01-100 mg/kg body weight/day of the inhibitor can be administered to a patient receiving these compositions.
It should also be understood that a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease being treated. The amount of inhibitor will also depend upon the particular compound in the composition.
According to another embodiment, the invention provides methods for treating or preventing a p38-mediated condition comprising the step of administering to a patient one of the above-described pharmaceutical compositions. The term xe2x80x9cpatientxe2x80x9d, as used herein, means an animal, preferably a human.
Preferably, that method is used to treat or prevent a condition selected from inflammatory diseases, autoimmune diseases, destructive bone disorders, proliferative disorders, infectious diseases, degenerative diseases, allergies, reperfusion/ischemia in stroke, heart attacks, angiogenic disorders, organ hypoxia, vascular hyperplasia, cardiac hypertrophy, and thrombin-induced platelet aggregation.
According to another embodiment, the inhibitors of this invention are used to treat or prevent an IL-1, IL-6, IL-8 or TNF-mediated disease or condition. Such conditions are described above.
Depending upon the particular p38-mediated condition to be treated or prevented, additional drugs, which are normally administered to treat or prevent that condition may be administered together with the inhibitors of this invention. For example, chemotherapeutic agents or other anti-proliferative agents may be combined with the p38 inhibitors of this invention to treat proliferative diseases.
Those additional agents may be administered separately, as part of a multiple dosage regimen, from the p38 inhibitor-containing composition. Alternatively, those agents may be part of a single dosage form, mixed together with the p38 inhibitor in a single composition.